螨变应原注射液体外效力评价方法的建立及验证

目的：针对铝佐剂吸附的螨变应原注射液缺乏有效的生物效力评价方法的现状，探索用基于ImmunoCAP仪的荧光酶联免疫分析方法（ImmunoCAP法）作为该制品体外效力评价方法可行性，以促进该制品质量标准的提高。方法：利用ImmunoCAP法测定螨变应原注射液对过敏病人血清中IgE的抑制率（又称为变应原活性），作为该制品效力评价的检测指标。研究中分析了该方法的特异性；探索了孵育时间、温度、血清稀释度、变应原浓度等条件对抑制率的影响；验证了该方法的精密性；测定了13批维持剂量螨变应原注射液的总变应原活性和上清游离变应原活性。结果：应用ImmuonCAP法测定维持剂量螨变应原注射液对过敏病人血清IgE抑制率，具有较好的特异性；维持剂量制品对血清中特异性IgE的抑制率与变应原浓度具有明显的量效反应关系；该法具有很好的精密性，RSD最大不超过5%；13批维持剂量制品总变应原对血清中IgE抑制率均大于50%；上清中游离变应原对血清中IgE抑制率均不高于20%。结论：ImmunoCAP法可用于铝佐剂吸附的螨变应原注射液的体外效力评价，重复性好，精密度高，操作简便。

Objective: To discuss the feasibility of a fluorescence ELISA method based on ImmunoCAP ^® for in vitro potency evaluation of mite allergen injection absorbed in view of the lack of effective evaluation method for the mite allergen injection adsorbed by aluminum adjuvant in order to improve the quality standard of the products. Methods: The inhibition rate of IgE in serum of allergic patients (also known as allergen activity) was determined by ImmunoCAP as the evaluation indicator of in vitro potency. The specificity of the method was analyzed. The effects of parameters such as incubation time, temperature, serum dilution and allergen concentration on inhibition rate were investigated. The precision of the method was verified. The total allergen activity and the free allergen activity of supernatant were determined in 13 batches of mite allergens injections of maintenance dose. Results: The specificity of the inhibition rate of IgE in serum of allergic patients of maintenance dose of mite allergen injection determined by ImmunoCAP was good. The inhibitory rate of the specific IgE in the serum of the maintenance dose preparation has a significant dose-response relationship with the allergen concentration. The method had good precision and the maximum RSD was no more than 5%. The inhibition rate of IgE in serum of the total allergen of 13 batches of maintenance dose was more than 50%. The inhibitory rate of IgE in serum of the free allergen in the supernatant was no more than 20%. Conclusion: The ImmunoCAP method can be used for in vitro potency evaluation of the mite allergen injection adsorbed by aluminum adjuvant,with good repeatability, high precision and easy operation.

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